LOCUS       sequence-364835-        6924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for melatonin in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  AUTHORS   Ino D, Tanaka Y, Hibino H, Nishiyama M
  TITLE     A fluorescent sensor for real-time measurement of extracellular 
            oxytocin dynamics in the brain.
  JOURNAL   Nat Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x.
  PUBMED    36138174
REFERENCE   2  (bases 1 to 6924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x."
COMMENT     Sequence Label: pCIS.MTRIA-MTN
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..581
                     /label=chicken beta-actin promoter
     intron          582..1598
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1522..1544
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1606..1625
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1654..1670
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1655..1671
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2558..2579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2685..2706)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3451..3467)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3451..3467)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3556..3575)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3621..3676
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3675..3694)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4031..4049)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4070..4086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4083..4105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4094..4110
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4118..4124,4125..4142,4143..4148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4163..4184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4301..4318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4472..5060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4552..4571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5231..6022,6023..6091))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5854..5873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6092..6196)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6223..6678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6310..6329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6520..6541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6804..6826
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6818..6835
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6819..6835
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6842..6861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6842..6860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg sggggggggg
      421 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      481 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      541 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      601 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      661 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      721 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      781 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
      841 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
      901 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
      961 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1021 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     1081 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     1141 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     1201 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     1261 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1321 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1381 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1441 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1501 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1561 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1621 ttgtgctgtc tcatcatttt ggcaaaggat tcctcgaggt cgacggtatc gataagcttg
     1681 atatcgaatt caccatgctt caaaatgggt ctcacctgaa cacctcctcg caggaccccg
     1741 tcctgggccg cccgccgtgg gtcaccacaa ctttagggtg cttcctcatc ttcactatcg
     1801 tcgtcgacat tcttggcaat ctgttggtca tcttctccgt gtaccggaac aaaaagctga
     1861 ggaacgcagg aaacatcttt gtggtgagcc ttgcagtggc agaccttgtg gtggctatct
     1921 atccataccc cctggtcctc acttccattt tcaacaatgg ctggaacctg ggatctgtcc
     1981 attgtcaaat cagcggcttc ctcatgggtg tcagcgtcat cggctccatc ttcaacatca
     2041 ccgccatcgc catcaacaga tactgctaca tatgtcacag cctcaaatac gacaagctgt
     2101 acagcgacaa aaactcagtg tgctatgtga ttttaatatg ggcgttgact gtggtggcca
     2161 tcgtacccaa cctgtttgtg ggttcactca agtatgaccc ccgagtttat tcctgcacct
     2221 tcgaacagtc ggccagctca gcgtatacaa tcgcagtggt gttttttcat tttattttac
     2281 ccatcatgat tgtcacctac tgctatttgc gcatttgcta caaaatatgg cgaaatttca
     2341 aagggaaatt aaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     2401 tccacatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     2461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcaga
     2521 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2581 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggatga
     2641 tggtgagaaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2701 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     2761 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2821 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2881 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     2941 tcaaggacga cggccactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3001 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3061 agctggagta caactcagta atctccaagg caaagatacg cacagtgaaa atgatgtttg
     3121 ttgtctttgt gctctttgct gtctgctggg cacccctcaa cttcatcgga ctggctgtag
     3181 caatcaaacc ggaggtggtg gttcccttaa tcccggagtg gttatttgtg gccagctact
     3241 tcatggccta cttcaacagc tgccttaacg ccattgtgta cggtgtgctg aaccagaact
     3301 tccggcggga gtacaaacgc atcgtagtgt ctgtgtgtac ggctcgcatc ttcttccagg
     3361 acagctccaa cgatgcaggg gagaggctca aaagtaaacc ctcaccgctc atgaccaaca
     3421 ataaccaggt caaagtggag tccgtctgag gatccactag ttctagagcg gccgcttcac
     3481 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     3541 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3601 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3661 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3721 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     3781 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     3841 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     3901 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     3961 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tggagctcca
     4021 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     4081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4201 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     6241 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6301 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6361 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6421 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6481 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6541 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6601 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6661 tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     6841 gtaatacgac tcactatagg gcgaattggg tacctagtaa tcaattacgg ggtcattagt
     6901 tcatagccca tatatggagt tccg
//
